5alpha-hydroxy-6beta-amino steroids and the preparation thereof



United States Patent 3,243,431 Sa-HYDROXY-tSfi-AMINO STERUIDS AND THE PREPARATION THEREOF Colin Leslie Hewett, Glasgow, and David Samuel Savage, Renfrewshire, Scotland, assignors to Organon Inc., West Orange, N.J., a corporation of New Jersey No Drawing. Filed July 23, 1964, Ser. No. 384,797 2 Claims. (Cl. 260239.5)

The invention relates to novel 6,8-amino steroids and to a process for the preparation thereof and to salts thereof. According to the invention there is provided a process which comprises reacting a oz,6a-oxido steroid of the androstane or pregnane series with a compound of the formula HNR R wherein R represents hydrogen, a lower alkyl group or a substituted lower alkyl group, R represents a lower alkyl group or a substituted lower alkyl group, or NR R represents a heterocyclic amino radical, to form the corresponding 5a-hydroxy-6fi-amino-steroid.

In particular, the invention relates to a process wherein a 5u,6a-oxido steroid having the formula:

in which X represents H(OH) or H(OAcyl); Y represents H(BOH), H(BOAcyl), =0, H(COCH H (CHOHCH or H (CHOAcylCH pound having the formula:

l l Y NRiRz in which X, Y, R and R have the meaning given above.

The invention further relates to novel 5a-hydroxy-6B- amino steroids of the pregnane and androstane series, with special reference to Gift-amino steroids having the formula:

in which R represents hydrogen, a lower alkyl group or a substituted lower alkyl group, R represents a lower alkyl group or a substituted lower alkyl group, or NR R represents a heterocyclic amino radical, X represents H(OH) or H(OAcyl) and Y represents (B (H Y a). H(CHOHCH or H(CHOAcylCH The 5a-hydroxy-6B-amino steroids, which are the new compounds of the invention, possess-important biological activities. They are central nervous system stimulants and depressant, possessing anaesthetic, sedative, anticholinergic and hypotensive properties.

The starting materials used in the proces herein decribed consist of 5a,6oc-oxido-steroids and, in particular, compounds having the formula:

wherein X and Y have the meanings given above.

The said 5a,6a-oxido steroids may be prepared by epoxidising the corresponding A steroid using, for example, an organic peracid such as perbenzoic acid or perphthalic acid.

The 5u,6a-oxido steroids are converted into the corresponding 5a-hydroxy-6fl-amino steroids by reacting with a compound of the general formula H.NR R in which R and R have the meanings given above. It has been found that the presence of water accelerates the reaction between the ,6a-0Xid0 steroid and the compound H.NR R If desired, the reaction may be carried outin the presence of a suitable organic solvent. The reaction is preferably carried out at the boiling point of the compound H.NR R or the solvent, if used, for some time, for example 50 to hours. If water is added to the reaction mixture the time may be reduced to 20 hours.

The following are examples of the compounds according to the formula H.N.R R which may be used in the process of the present invention; methylamine, ethylamine, propylamine, butylamine, dimethylamine, diethylamine, morpholine, piperidine, piperazine and pyrrolidine.

Starting with the 3-, 17- or 20-acyloxy-5a,6a-oxidosteroids there is obtained a mixture of the corresponding 3-, 17- or 20-acyloxy and free hydroxy-5a-hydroxy- 6,8-arnino steroid which are readily separable. The degree of hydrolysis of the acyloxy groups depends on the reaction time and the amount of water present. The acyloxy compounds may be hydrolysed to the corresponding free hydroxy compounds with alkali in a known manner, likewise the free hydroxy groups, except the SOL-hydroxy group, may be esterified in the usual way.

Where the starting 5a,6oz-OXidO steroid contains a keto group this may be converted into a hydroxyl group by reduction either in the starting oxide or in the Sa-hydroxy 6,8-amino steroid compound prepared from the keto-oxide. For example, the 5a-hydroxy-6p-amino-l7- keto-steroids may be converted to the corresponding 5a- 17fl-dihydroxy-6B-amino steroids by reduction.

As reducing agents may be used, for example, sodium borohydride in methanol. In a similar manner, the 20- ketone in a a-hydroxy-6fi-amino-20-kto-steroid may be reduced to a 20-hydr-oxy group. When sodium or potassium borohydn'de is used for this reduction any 3- acyloxy group remains unchanged and esterification will yield mixed esters.

Any secondary hydroxy group present in the SOL-hydroxy-6/3-amino steroids or any secondary acyloxy group after saponification, for example, the 3-hydroxy, 17-hydroxy and 20-hydroxy may be converted into the corresponding ketone by oxidation, for example, according to the Oppenauer method or by means of chromium trioxide.

The 5a-hydroxy-6fi-amino-steroids according to the present invention may be converted into salts thereof, for example, organic salts such as citrates, or pyruvates or inorganic salts such as the hydrochlorides or they may be converted into the quaternary ammonium salts with alkylhalides.

The invention is illustrated further by the following examples.

Example I A solution of 5a,6a-epoxy-pregnan-3fi-ol-ZO-one (5.6 gm.) in morpholine (15 ml.) was refluxed for 5 days, the solution cooled and the product precipitated by the addition of water. The white solid was filtered off, washed Well with water, dried and suspended in boiling ether (300 ml.) for 5 minutes, cooled and the sparingly soluble crystalline :solid filtered off (3.6 gm.) M.P. 255- 265. Recrystallisation from acetone gave 6,8-morpholino 5a-pregnan-3 8,5a-diol-20-one as small prisms, M.P. 276-280 (dec.), [111 -1l.4 (c. 0.2). The 3-acetate, prepared from the diol (500 mg.) in acetic anhydride (4 ml.) and pyridine (4 ml.) at 100 for 6 hours, crystallised from ether in small prisms, M.P. 228230 (dec.). Recrystallisation from acetone-ether gave prisms, M.P. 228230 (dec.), [ab -12 (0. 0.4).

In the same manner other 3-esters have been prepared derived from valeric acid, capric acid, lauric acid and fl-phenyl propionic acid.

Hydrolysis of the 3,B-mono-acetate (200 mg.) with potassium carbonate (250 mg.) in water (2.5 ml.) and methanol (20 ml.) at the boil for 40 minutes yielded the original diol, M.P. 272-276.

Example 11 A solution of 5a,6a-epoxy-pregnan-3fi-ol-20-one 3- acetate (20 gm.) in morpholine (50 ml.) was refluxed for days; the solution was cooled and Water added to precipitate a sticky solid which gradually hardened on-stirring to a white solid. After Washing well with water the product was crystallised from acetone to give several crops (total 12.2 gm.) consisting essentially of crude diol, M.P. 202270 (dec.).

The material obtained from the acetone mother liquors after filtering through alumina in ethereal solution, gave the 3-acetate (4.5 gm.), M.P. 224-228 (dec.).

The crude diol (12.2 gm.) in methanol (350 ml.) was boiled under reflux for 1 hour with potassium carbonate (10 gm.) in water (12 ml.). After addition of water the pure 6B-morpholino-5a-pregnan-3,8,5a-diol-20-one was filtered off and dried (9.2 gm.), M.P. 274278 (dec.).

Example 111 A solution of 5a,6a-epoxy-pregnan-3,8-ol-20-one 3- acetate (5 g.) in morpholine (20 ml.) and Water (10 ml.) was boiled under reflux for 20 hours. Addition of water to the cooled solution precipitated the product which was filtered, washed with water and hydrolysed with 4% aqueous methanolic potassium hydroxide solution. The suspension was concentrated and the sparingy soluble p oliuo i l (4. a). M.P. 2.702.77 (dec.). filtered off.

4 Example IV Sodium bore-hydride (1 g.) was added over 20 minutes to a solution of 6fl-morpholino-5a-pregnan-35,5a-di0l-20- one Fifi-acetate (2.8 g.) in methanol ml.) and the solution allowed to stand at 20 for 1 hour. A crystalline material had precipitated out and addition of water yielded a crystalline product which was recrystallised from acetone in small prisms (1.85 g.) of 6/3-morpholino-5a-pregnan- 3fi,5a,20,6-triol 3fl-acetate, M.P. 268270, [111 70.1" (c. 0.5). 1

A solution of potassium carbonate (3 g.) in water (15 ml.) was added to a solution of the triol-monoacetate (1.8 g.) in methanol (50 ml.) and the mixture refluxed for 1 hour. The product was precipitated from the cooled solution by addition of water, filtered off and crystallised from acetone yielding the triol in small needles (1.4 g.), M.P. 234-238 (dec.), [11],; 61.6 (c. 1.0).

Example V A solution of 313,20,8-diacetoxy-5a,GwepOXy-pregnane (7.9 g.) in morpholine (34 ml.) and water (8 ml.) was boiled under reflux for 42 hours, concentrated almost to dryness, and water added to precipitate a buff-coloured solid which was filtered oif and well washed with water.

This dried solid was dissolved in ether (50 ml.) and the solution filtered down a column (6 x 1 /2" dia.) of alumina. Elution with ether ml.) yielded a clear gum which was dissolved in ether, light petrol (B.P. 40/60) added and the ether distilled off precipitating a crystalline solid (3.4 g.), M.P. 206-208"; recrystallisation from ether yielded 3B,20 3-diacetoxy-6 3-morpholino-5w pregnan-5a-ol in heavy prisms, M.P. 206208, [04] 37.

Further elution with ether (1.5 1.) yielded fractions (2.8 g.), melting range 230-244. Recrystallisation from ether yielded 20/3-acetoxy-6fl-morpholino-5u-pregnan-3/3, 5 a-diol in fine needles, M.P. 243-245 Elution with methanol (250 ml.) and evaporation of the solvent gave a brown crystalline solid which was recrystallised from acetone to give 6 8-morpholino-5oz-pregnan- 3[3,5oz,20fl-tl'i0l in small needles (250 mg), M.P. 235-239 (dec.); the melting point was undepressed on admixture with an authentic specimen.

Example VI A solution of 5a,6a-epoxy-pregnan-3B-ol-20-one 3B- acetate (10 g.) in water piperidine was refluxed for 70 hours (1:1, 60 ml.). The product, precipitated as a yellow gum by the addition of water to the cooled reactant solution, was extracted with ether, the extract dried (NaSO and the solution evaporated to dryness. The residual yellow gum was hydrolysed in the usual manner, in aqueous methanolic potassium carbonate solution, the product extracted with ether and concentrated to give a crop of fine needles (5.3 g.), M.P. 170174 (dec.). Recrystallisation from ether gave the piperidino-diol in fine needles, M.P. 170174 (dec.).

Perchloric acid (0.1 ml.) was added to a solution of the piperidino-diol (2.75 g.) in acetic anhydride (2 ml.), acetic acid (15 ml.) and 30% hydrogen bromide; acetic acid (2 ml.) and the solution allowed to stand at 20 for 30 minutes. Careful addition of 50% aqueous potassium hydroxide solution precipitated a solid which was filtered off and recrystallised from ether to give needles, M.P. 188190, softening ca Recrystallisation from ether gave the monoacetate in needles, M.P. 188190, [a1 -21.5 (c. 2).

In the same way this compound has been converted into the 3/3-acylates derived from butyric acid, oenanthic; acid, ,B-phenyl propionic acid, and succinic acid.

Example VII A suspension of 5a,6a-epoxy-pregnan-3/8-ol-20-one (10 g.) in piperazine-hexahydrate was refluxed for 72 hours. Water (30 ml.) was added to the cooled mixture and the resulting suspension filtered. The insoluble white solid was well washed with water, dried and suspended in boiling acetone. A sparingly soluble white crystalline solid (8 g.), M.P. 250260 (dec.) was filtered oif. Recrystallisation from methanol-acetone yielded the piperazino-diol in plates, M.P. 260275 (dec.), [od 9 (c. 0.3).

Example VIII A solution of 5a,6a-epoxy-pregnan-3B-ol-20-one 3B- acetate (10 gm.) in waterzbutylamine (1:3, 40 ml.) was refluxed for 70 hours. The solution was evaporated to dryness under reduced pressure and the residual yellow gum hydrolysed in the usual manner in aqueous methanolic potassium carbonate solution. The product was dissolved in benzene, filtered down a column (3" X 1" dia.) of alumina, and eluted with benzene (200 ml.). Evaporation of the benzene and crystallisation of the residual yellow gum from ether gave small prisms (7 g.), M.P. 198-202. Recrystallisation from ether gave 6/3- butylamino-5a-pregnan-3fl,5o-diol-20-one in prisms, M.P. 198-202", +24 (c. 2). In the same way, using methylamine, propylamine and diethylamine the above-mentioned a,6oc-epoxy-compound has been converted into the corresponding Sa-hydroxy- 6fl-amino-derivatives.

Example IX A solution of 5a,6a-epoxy-androstan-3fi-ol-17-one 3,8- acetate (20 g.) in morpholine (40 ml.) was refluxed for 10 days, the solution cooled, and water added to precipitate a yellow solid which was taken up in methanol (150 ml.), a solution of potassium carbonate (10 g.) in water (30 ml.) added, and the solution refluxed for 1 hour. The suspension was cooled and the sparingly soluble crystalline material (12.3 g.), M.P. 268280 filtered off and washed with ice-cold methanol (10 ml.).

Recrystallisation of this material from acetone gave 6B-morpholino-Sa-androstan-3fl,5a-diol-17-one in needles, M.P. 282286 (dec.), [a] 9 (c. 0.3).

A suspension of the morpholino-diol (2 g.) in pyridine (14 ml.) and acetic anhydride (14 ml.) was heated into solution at 100 for 1 hour. The product, precipitated from the cooled solution by the addition of water, was extracted with ether, the extract well washed with water and dried (NaSO The concentrated extract was filtered down a column (3" X 1") of alumina and the ether eluate (500 ml.) concentrated and crystallisation efl'ected from acetone to give 6,8-morpholino-5a-androstan-3B,5u-diol- 17-one 3fl-acetate as large prisms, M.P. 177179, [a] 14.8 (c. 0.4).

In an analogous manner the 3/8-acylates derived from propionic acid, caproic acid, fi-phenyl propionic acid and palmitic acid have been prepared.

Example X Sodium borohydride (0.25 g.) was added over 15 minutes to a stirred solution of 6,8-morpholino-5u-androstan-3fl-5a-diol-17-one (1 g.) in methanol (60 ml.) and stirring continued for 2 hours. Addition of a 5% solution of potassium hydroxide precipitated a white solid which was filtered off and crystallised from acetone to give 6,8-morpholino-5a-androstan-3 8,5a-17fi-triol in small needles (700 mg), M.P. 233-236", [u] -58.6 (c. 1).

Example XI Soduim borohydride (1 g.) was added over 20 minutes to a stirred solution of 6B-morpholino-5u-androstan-3B, Soc-diOl-l'I-Olle 3,8-acetate (3 g.) in methanol (20 ml.) and stirring continued for 1 hour at 20. Addition of water precipitated the product as a white crystalline solid (2.73 g.), which was filtered off and recrystallised from aqueous acetone to give the triol-monoacetate, in small needles, M.P. 185-189, [0th, 70 (c. 2).

6 Example XII A solution of 3B,17/3-diacetoxy-5a,6a-epoxy-androstane (10 g.) in morpholine (32 ml.) and water (8 ml.) was boiled under reflux for 66 hours, and the solution evaporated to dryness under reduced pressure to give a brown gum which gave a light brown crystalline solid on stirring with water. This solid was filtered ofl, dried at 47 under reduced pressure, and finely ground before extracting well with either. The sparingly soluble material (4.5 g.) was dissolved in acetone, boiled with charcoal, filtered and concentrated to give 6/i-morpholino-5u-androstan-3,8, 5a,17,B-triol in small needles, M.P. 235238.

The ether mother liquors were concentrated (to 50 ml.) and filtered down a column (6" x 1") of alumina. Elution with ether (70 ml.) gave a fraction (3 g.), M.P. 138- 143. Recrystallisation from ether gave 35,17B-diacetoxy-6fl-morpholino-5a-androstan5a-ol in small needles, M.P. 145-149, [01],; 68 (c. 2).

Further elution with ether and recrystallisation from ether gave l7B-acetoxy-6B-morpholino-5a-androstan-3B, SOL-diol. in micro-needles, M.P. 238240, [aJ 58 (c. 0.8).

Example XIII A suspension of 5a,6ot-epoxy-androstan-313-01-17-one 3fl-aoetate (10 g.) in piperidine2water (3:1, 40 ml.) was refluxed for 22 hours. Addition of water precipitated a gum which was taken up in 3% aqueous methanolic potassium hydroxide solution (70 ml.) and refluxed for 1 hour. A heavy crystalline solid (9 g.) deposited and was recrystallised from acetone in needles, M.P. 210- 212. Recrystallisation from ether gave the piperidinodiol in small needles, M.P. 205-207, [ab +3.5 (c. 2).

In an analogous manner 5w6a-epoxy-androstan-35-01- 17-one 3B-acetate has been converted into the corresponding 3,8,5a-dihydroxy-6B-amino compounds, of which the amino group is derived from ethylamine, dimethylamine and propylamine.

The piperidino-diol (5.4 g.) was acetylated in the usual manner with acetic anhydride, 30% hydrogen bromidezacetic acid, and a catalytic amount of perchloric acid. crystallisation from ether gave the 3B-monoacetate in rods, M.P. 123, [ed 17 (0. 2).

In an analogous way the 3,8-mono acylates derived from trimethyl acetic acid, oenanthic acid, capric acid, fi-phenyl propionic acid and stearic acid have been pre pared.

We claim:

1. Steroids of the formula:

OH NRi z in which X is selected from the group consisting of H(OH) and H(OAcyl),

R and R are selected from the group consisting of hydrogen, a lower alkyl group, and a substituted lower alkyl group, R and R not being hydrogen simultaneously, and when taken together form with the nitrogen atom a heterocyclic ring selected from the group consisting of morpholine, piperidine, piperazine and pyrrolidine, and

R is selected from the group consisting of O, H(OH) and H(OAcyl), in which the acyl group is derived from an organic carboxylic acid having 1-18 carbon atoms.

2. Salts of the compounds of claim 1 wherein the salt is selected from the group consisting of quaternary am- 8 monium salts and the aciZ-addition salts derived from OTHER REFERENCES organic and inorganic acids Batres et al.: J. Org. Chem., vol. 26, pp. 878-80 References Cited by the Examiner UNITED STATES PATENTS 5 LEWIS GOTTS, Primary Examiner. 3123623 3/1946 sassaki 260.3973 HENRY A. FRENCH, Assistant Examiner,

3,156,710 11/1964 Sassaki 260-349 

1. STEROIDS OF THE FORMULA: 